目的 探究黃芩莖葉總黃酮（SSTF）治療潰瘍性結(jié)腸炎（UC）合并貧血的藥效及作用機(jī)制。方法 通過(guò)文獻(xiàn)查閱和Swiss Target Prediction數(shù)據(jù)庫(kù)獲取SSTF中黃酮成分及其作用靶點(diǎn)；通過(guò)GeneCards、OMIM和TTD數(shù)據(jù)庫(kù)收集UC合并貧血疾病靶點(diǎn)；使用Venny 2.1繪圖軟件獲取藥物和疾病交集靶點(diǎn)；將交集靶點(diǎn)導(dǎo)入STRING數(shù)據(jù)庫(kù)構(gòu)建蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)；采用Metascape數(shù)據(jù)平臺(tái)進(jìn)行基因本體（GO）功能和京都基因和基因組百科全書（KEGG）通路富集分析；再采用AutoDock 1.5.7軟件對(duì)主要活性成分和關(guān)鍵靶點(diǎn)進(jìn)行分子對(duì)接。利用噁唑酮建立UC小鼠模型，模型成功小鼠隨機(jī)分為模型組、溶媒（0.5%羧甲基纖維素鈉）組、柳氮磺吡啶腸溶片（SASP，陽(yáng)性藥，450 mg·kg⁻¹）組和SSTF低、中、高劑量（26、52、104 mg·kg⁻¹）組，每組15只。另設(shè)15只小鼠作為對(duì)照組，不造模。各治療組從第9天開始ig相應(yīng)藥物，對(duì)照組與模型組給予0.9%的氯化鈉溶液，每天1次，連續(xù)7 d。從第8天開始每天稱小鼠體質(zhì)量，進(jìn)行疾病活動(dòng)指數(shù)（DAI）評(píng)分；給藥結(jié)束后取材，測(cè)量結(jié)腸長(zhǎng)度，對(duì)脾臟組織稱質(zhì)量，并計(jì)算脾臟指數(shù)；蘇木素-伊紅（HE）染色進(jìn)行結(jié)腸組織病理學(xué)檢查，酶聯(lián)免疫吸附試驗(yàn)（ELISA）試劑盒法檢測(cè)血清腫瘤壞死因子α（TNF-α）、白細(xì)胞介素10（IL-10）、促紅細(xì)胞生成素（EPO）、血紅蛋白（HGB）、鐵（SI）、鐵蛋白（SF）、鐵調(diào)素（Hepcidin）和可溶性轉(zhuǎn)鐵蛋白受體（sTfR）水平，實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測(cè)脾組織促紅細(xì)胞生成素受體（EPOR）、赤鐵酮（ERFE）、膜鐵轉(zhuǎn)運(yùn)蛋白（FPN/SLC40A1）和肝組織鐵調(diào)素（Hepcidin）、FPN mRNA表達(dá)。結(jié)果 網(wǎng)絡(luò)藥理學(xué)結(jié)果表明，SSTF中27個(gè)黃酮成分可作用于EPO、EPOR等25個(gè)交集靶點(diǎn)，調(diào)控JAK/STAT等信號(hào)通路。分子對(duì)接結(jié)果表明主要活性成分黃芩苷、漢黃芩苷、木犀草素、野黃芩苷、野黃芩素、芹菜素與關(guān)鍵靶點(diǎn)間結(jié)合穩(wěn)定。體內(nèi)實(shí)驗(yàn)結(jié)果表明，與模型組比較，SSTF組小鼠體質(zhì)量顯著升高、DAI評(píng)分顯著降低、脾臟指數(shù)顯著降低、結(jié)腸長(zhǎng)度顯著增加（P＜0.05、0.01）；結(jié)腸組織病理?yè)p傷明顯改善；血清中Hepcidin、sTfR、TNF-α水平顯著降低（P＜0.05、0.01），血清SF、SI、HGB、EPO、IL-10水平顯著升高（P＜0.05、0.01）；小鼠脾組織中EPOR、ERFE、FPN mRNA表達(dá)水平和肝組織中FPN mRNA表達(dá)水平顯著升高（P＜0.05、0.01），肝組織中Hepcidin mRNA表達(dá)水平顯著降低（P＜0.05）。結(jié)論 SSTF可通過(guò)調(diào)節(jié)EPOR/ERFE/FPN信號(hào)軸治療UC合并貧血。

Objective To explore the efficacy and mechanism of action of Scutellaria Baicalensis stem-leaf total flavonoids (SSTF) in the treatment of ulcerative colitis (UC) complicated with anemia. Methods The flavonoid components and their target sites in the stems and leaves of Scutellaria baicalensis were obtained through literature review and the Swiss Target Prediction database. The target sites of ulcerative colitis (UC) combined with anemia were collected from the GeneCards, OMIM and TTD databases. The Venny 2.1 drawing software was used to obtain the intersection target sites of drugs and diseases. The intersection target sites were imported into the STRING database to construct a protein-protein interaction (PPI) network. The Metascape data platform was used for gene ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. The AutoDock 1.5.7 software was used for molecular docking of the main active components and key target sites. The UC mouse model was established by using oxazolone. The successfully modeled mice were randomly divided into the model group, the solvent (0.5% carboxymethyl cellulose sodium) group, the sulfasalazine enteric-coated tablet (SASP, positive drug, 450 mg·kg⁻¹) group and the SSTF low, medium and high dose (26, 52, 104 mg·kg⁻¹) groups, with 15 mice in each group. Another 15 mice were set as the control group without modeling. The treatment groups were given the corresponding drugs by intragastric administration from the 9th day, while the control group and the model group were given 0.9% sodium chloride solution once a day for seven consecutive days. The body weight of the mice was measured every day from the 8th day, and the disease activity index (DAI) score was evaluated. After drug administration, the colon length was measured, the spleen tissue was weighed and the spleen index was calculated. The colon tissue was stained with hematoxylin-eosin (HE) for histopathological examination. The levels of serum tumor necrosis factor α (TNF-α), interleukin 10 (IL-10), erythropoietin (EPO), iron (SI), ferritin (SF), hepcidin and soluble transferrin receptor (sTfR) were detected by enzyme-linked immunosorbent assay (ELISA) kits. The mRNA expressions of erythropoietin receptor (EPOR), erythroferrone (ERFE), ferroportin (FPN/SLC40A1) in spleen tissue and hepcidin, FPN and Hepcidin in liver tissue were detected by real-time fluorescence quantitative PCR (qRT-PCR). Results The results of network pharmacology indicated that 27 flavonoid components in SSTF could act on 25 intersection targets such as EPO and EPOR, and regulate signaling pathways such as JAK/STAT. The results of molecular docking showed that the main active components, baicalin, wogonin, luteolin, scutellarein, and apigenin, had stable binding with key targets. The in vivo experiments demonstrated that compared with the model group, the body weight of mice in the SSTF group significantly increased, the DAI score significantly decreased, the spleen index significantly decreased, and the colon length significantly increased (P < 0.05, 0.01); the pathological damage of colon tissue was significantly improved; the levels of Hepcidin, sTfR, and TNF-α in serum significantly decreased (P < 0.05, 0.01), while the levels of SF, SI, HGB, EPO, and IL-10 in serum significantly increased (P < 0.05, 0.01); the mRNA expression levels of EPOR, ERFE, and FPN in spleen tissue and FPN in liver tissue significantly increased (P < 0.05, 0.01), and the mRNA expression level of Hepcidin in liver tissue significantly decreased (P < 0.05). Conclusion SSTF can treat UC combined with anemia by regulating the EPOR/ERFE/FPN signaling axis.

R285.5

泰州市“鳳城英才計(jì)劃”青年科技人才托舉工程（TZ-018）